NECi, The Nitrate Elimination Co., Inc., develops and commercializes enzyme-based products for testing and remediation of compounds of environmental concern. NECi focuses on the application of higher plant Nitrate Reductase (NaR) to problems caused by excess nitrate in the environment, a pollution and human health issue worldwide. NECi proposes to apply company experience with electrically-driven enzyme systems to development of an amperometric NaR-based Nitrate Biosensor (NaR-NBS). NECi proposes to determine whether NaR, a redox enzyme with three internal electron-carrying cofactors, can function when bonded directly to electrode surfaces (electropolymers, noble metals, etc.). Alternatively, NECi will co-immobilize NaR and an electron-carrying dye onto inert particles, and embed these particles in electrode materials such as carbon paste. Either configuration will result in a "reagent-less" electrode for incorporation into an NaR-based Nitrate Biosensor (NaR- NBS). The working hypothesis is that NaR electrodes will effectively reduce nitrate in proportion to current used, and can be calibrated using nitrate standards. Prototypes incorporating both natural (corn seedling) and Pichia-expressed NaR will be performance-tested using standards and real world samples for sensitivity, reaction time, and stability. In Phase II, the most promising NaR/electrode material prototypes will be optimized, and production techniques and sensor electronics package will be developed. PROPOSED COMMERCIAL APPLICATIONS: The market for water testing equipment is US$ billions over the next ten years. There is a need for new technology for nitrate monitoring such as the NaR-NBS. Improved nitrate detection products will find many applications in monitoring of environmental quality; in testing of potable water in contaminated areas; monitoring industrial process and effluent waters; septic tank monitors, and in vivo monitoring for gauging NO levels. The NECi Nitrate Biosensor will be an electronic device with replaceable enzyme cartridges. Generic enzyme electrode technology will be adapted for Nitrite, Nitrous Oxide, and Nitric Oxide Biosensors for a suite of implantable sensors.